타우린이 고온에서 육계 배아 pectoralis의 배양 시 protein turnover에 미치는 영향 = Effect of taurine on protein turnover in embryonic pectoralis of broiler cultivated at high temperatures.

다국어 초록 (Multilingual Abstract)

The study was investigated the effects of the taurine on protein turnover in embryonic pectoralis of broiler cultivated at high temperature. This study consisted of three trials. The first trial was to investigate the effects of taurine on RNA synthesis in nuclei isolated from chick embryo pectoralis at various temperatures. The second trial was to testify the effects of taurine on in vitro protein synthesis and protein degradation in the cell free system, such as pectoralis homogenates of broiler chick embryos at various temperatures. The third trial was to investigate the effects of taurine on in vitro protein synthesis and protein degradation and HSP 70 concentrations in the pectoralis slices of broiler chick embryos at various temperatures. In the first trial, RNA synthesis in nuclei decreased at high temperature but taurine had no effect on RNA synthesis in nuclei at various temperatures. The result of first trial suggested that the effect of taurine on RNA synthesis in nuclei at various temperatures was concentration-dependent. In the second trial, pectoralis homogenate of chick embryos incubated at 37, 41, 45, 49℃ to prove the effect of high temperature on the protein synthesis and protein degradation. The higher temperature went up, the more the degradation rates of cytoplasmic protein in pectoralis homogenate of chick embryos increased. After SDS-PAGE, five-bands (190, 70, 60, 42, 24 kDa) were analyzed by LC-MS/MS. These proteins were identified as myosin heavy chain 1C, ovotransferrin BB type, serum albumin precursor, betaine-homocysteine S-methyltransferase 1 and triose Phosphate Isomerase. Among the bands, 70 and 60 kDa protein concentration was not significantly different, but other bands were decreased. Protein synthesis rate increased more time but were not affected by incubating temperatures. In the third trial, pectoralis slice of chick embryos incubated at 37, 41, 45, 49℃ to prove the effect of high temperature on the protein synthesis and protein degradation and HSP 70 concentrations. The higher temperature went up, the more the degradation rates of cytoplasmic protein in pectoralis homogenate of chick embryos increased. The result of third trial suggested that the effect of taurine was dependent on temperatures. In conclusion, heat stress leads to decrease protein synthesis and increase protein degradation in chick embryo pectoralis, and the effect of taurine on protein turnover in embryonic pectoralis was dependent on temperature and concentrations.

목차 (Table of Contents)

• Ⅰ. 서 론 1
• Ⅱ. 연구사 3
• 1. 고온 스트레스 시 육계의 생리적 변화 3
• 2. 타우린의 생리적 기능과 육계에 미치는 영향 4
• 2.1 타우린의 생리적 기능 4
• 2.2 타우린이 가금에 미치는 영향 5
• 3. 고온 스트레스 시 RNA합성에 대한 연구 5
• 4. 고온 스트레스 시 protein turnover에 대한 연구 6
• 5. HSP 70 에 대한 연구 6
• Ⅲ. 재료 및 방법 8
• 1. 공시동물 8
• 2. 육계 배아의 pectoralis로부터 추출한 핵에서 RNA 합성량 측정 8
• 2.1 육계 배아 pectoralis로부터 핵의 추출 8
• 2.2 핵으로부터 RNA 합성량 조사 8
• 3. 육계 배아 pectoralis의 homogenate에서 단백질 합성량 측정 9
• 3.1 육계 배아 pectoralis의 균질화 9
• 3.2 DC Protein assay kit에 의한 육계 배아의 pectoralis의 homogenate로부터 단백질량 측정 9
• 3.3 전기영동을 통한 육계 배아 pectoralis의 homogenate로부터 단백질량 분석 10
• 3.4 육계 배아 pectoralis의 homogenate로부터 단백질 합성 10
• 4. 육계 배아의 pectoralis slice에서 단백질의 양과 단백질 합성량 측정 11
• 4.1 배아 pectoralis slice 준비 11
• 4.2 육계 배아의 pectoralis slice로부터 단백질량 측정 11
• 4.3 육계 배아 pectoralis slice로부터 단백질합성 12
• 4.4 전기영동 및 Radiography 12
• 4.5 LC-MS/MS 13
• 4.5.1 In gel digestion 13
• 4.5.2 Identification of proteins by LC-MS/MS 13
• 4.6 Western blot 14
• 5. 통계 분석 15
• Ⅳ. 결과 및 고찰 16
• 1. 온도 처리 수준에 따라 타우린이 RNA 합성량에 미치는 영향 16
• 2. 육계 배아 pectoralis homogenate로부터 단백질의 양과 단백질 합성량 측정 19
• 3. 육계 배아 pectoralis slice로부터 단백질의 양과 합성량 측정 27
• 4. Western blot을 통한 HSP70 측정 39
• Ⅴ. 적요 41
• Ⅵ. 참고문헌 43